Application of Compounds in Anti-Dengue and Zika Virus Infection
A virus infection, dengue virus technology, applied in the field of compounds
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Embodiment 1
[0037] Embodiment 1, the preparation of compound 1-94
[0038]
[0039] Get intermediate A1 in 100ml round bottom flask, add 198mg Pd(dppf) 2 .DCM, 2g potassium phosphate and 1.2g p-trifluoromethylthiophenylboronic acid pinacol ester, add 10ml toluene, under the protection of argon, react at 100°C for 12h, spin the solvent, add 100ml water, extract with 20ml dichloromethane Twice, using petroleum ether: ethyl acetate = 30:1 to pass through a silica gel column to obtain 1 g of intermediate A3 (colorless liquid), with a yield of 64%.
[0040] Take 1g of intermediate A3 and 650mg of intermediate A4 in a 25ml round bottom flask, add 240mg of p-toluenesulfonic acid monohydrate and 10ml of n-butanol, react at 130°C for 16h, spin the solvent under reduced pressure, add water, and use 20ml of ethyl acetate Extracted twice, and passed through a silica gel column with ethyl acetate:petroleum ether=1:1 to obtain 320 mg of compound 1 (white solid), with a total yield of 24%.
[0041]...
Embodiment 2
[0044] Embodiment 2, the inhibitory activity of compound to dengue virus and Zika virus
[0045] Reagents and materials used: DMEM medium (Thermo), phenol red-free DMEM medium (Thermo), FBS (Thermo), PBS (Thermo), Antibiotic-antimycotic (Anti-anti, Thermo), 0.25% trypsin (Thermo) , DMSO (Sigma), MTT (Sigma), mineral oil (Sigma), 2-propanol (Sigma), absolute ethanol (Sigma), multi-source RNA extraction kit (Corning), cDNA synthesis kit (Bio -Rad), SYBR Green method Q-PCR master mix (Bio-Rad). 6-well, 48-well cell culture plate (Corning), 96-well low adsorption flat bottom cell culture plate (Corning), 96-well transparent V-bottom QPCR plate, 75cm 2 、25cm 2 Cell culture square bottle (Corning), 15\50ml centrifuge tube (Corning), 1.5ml centrifuge tube (Corning), 0.2ml 8-strip PCR tube (Corning), 5\10ml pipette (Corning).
[0046] Instruments used: multifunctional microplate reader (SkanIt, Thermo); optical microscope (Olympus); centrifuge (Centrifuge 5424, Eppendorf); sample g...
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